CN108434091B

CN108434091B - Self-healing hydrogel for promoting wound healing and treating tumors and preparation method thereof

Info

Publication number: CN108434091B
Application number: CN201810608139.2A
Authority: CN
Inventors: 雷波; 周丽; 薛语萌
Original assignee: Xian Jiaotong University
Current assignee: Xian Jiaotong University
Priority date: 2018-06-13
Filing date: 2018-06-13
Publication date: 2021-02-09
Anticipated expiration: 2038-06-13
Also published as: CN108434091A

Abstract

The invention relates to a self-healing hydrogel for promoting wound healing and tumor treatment and a preparation method thereof, wherein the self-healing hydrogel is a polyether-cationic polymer aqueous solution with the mass concentration of 8-50%, a polyether-p-hydroxybenzaldehyde aqueous solution with the mass concentration of 10-40% and a dopamine-modified bioactive inorganic material aqueous solution with the mass concentration of 2-10%, and the contents are as follows (22-28): (7-10): (4-10) mixing FCB hydrogel obtained by Schiff base reaction in a volume ratio, wherein F is polyether-cationic polymer; c is polyether-p-hydroxybenzaldehyde; b is a dopamine modified bioactive inorganic material. The compound has good biocompatibility, good photo-thermal property and strong antibacterial property, can effectively inhibit the growth of animal tumors under the irradiation of laser, and can promote the repair of the skin injury of mice. The preparation method is simple, has no organic solvent residue, and has the advantages of environmental protection, convenient operation and low raw material cost.

Description

Self-healing hydrogel for promoting wound healing and treating tumors and preparation method thereof

Technical Field

The invention relates to the technical field of biomedical materials, in particular to self-healing hydrogel for promoting wound healing and tumor treatment and a preparation method thereof.

Background

The skin, the largest organ of the human body, plays an important role in stabilizing the internal environment of the body and also protects the body from the external environment. Skin cancer is one of the common malignant tumors in humans, and more than 100 ten thousand cases of skin cancer are detected each year. Current clinical treatment strategies for skin cancer include surgical resection and late-stage chemotherapy/radiotherapy. In order to prevent recurrence, many normal skins around tumor cells must be removed during surgical excision, thereby causing large skin defects, easily causing wound infection, and leading to difficult wound healing. In addition, traditional chemotherapy/radiotherapy has been widely used to avoid cancer recurrence, but its serious side effects and drug resistance cause endless pain to patients. To date, meeting the three major goals of skin cancer treatment, skin repair after resection surgery, and anti-infection remains a significant challenge. Therefore, the development of materials for tumor treatment, promotion of wound repair and infection resistance has become a research hotspot in the technical field of biomedical materials.

In recent years, photothermal therapy has become a novel strategy in tumor therapy due to its high efficacy and low toxicity. Various photothermal agents such as: graphene oxide nanomaterials, gold nanoparticles, molybdenum disulfide, cuprous sulfide, and the like have been developed in succession for tumor therapy. Dopamine (DA) is widely used in the field of biomaterials due to its good biocompatibility and low toxicity. In addition, dopamine monomers can be self-polymerized into Polydopamine (PDA) with better photothermal properties. Under the alkalescent condition, PDA can be modified on the surface of the nano-particles through amidation reaction or Schiff base reaction, thereby endowing the nano-particles with photo-thermal performance. Moreover, due to the sensitivity of PDA to pH, the PDA modified nanoparticles can be stable under physiological conditions and degraded in the weakly acidic environment of tumors.

Bioactive Glass (BG) is a silicate glass based on a glass network formed by three-dimensional silica tetrahedra, incorporating Ca and P as modifiers. Bioceramics refer to a class of ceramic materials that serve a specific biological or physiological function such as: hydroxyapatite (HAP), Calcium Phosphate (CP), β -tricalcium phosphate (β -TCP), calcium phosphate bi-directional (BCP), Calcium Silicate (CS), Calcium Carbonate (CC), and the like. The bioactive glass and the biological ceramic are widely applied to the aspects of bone defect, skin tissue repair and reconstruction and the like due to good bioactivity, biocompatibility and excellent affinity performance with biological tissues to form chemical bonding with bone tissues and soft tissues and promote tissue regeneration.

However, the prior art also has the problems of weak cancer cell inhibition ability, poor anti-infection ability, susceptibility to infection of skin wound, poor healing and the like when the biological material is applied to skin cancer treatment.

Disclosure of Invention

Aiming at the problems in the prior art, the invention provides the self-healing hydrogel for promoting wound healing and tumor treatment and the preparation method thereof, the method and the process are simple, the prepared hydrogel has better photo-thermal property and stronger antibacterial property, can effectively inhibit the growth of tumor and promote the repair of skin wound, and has great application value in the aspect of skin cancer treatment.

The invention is realized by the following technical scheme:

a self-healing hydrogel for promoting wound healing and treating tumors is a polyether-cationic polymer aqueous solution with a mass concentration of 8% -50%, a polyether-p-hydroxybenzaldehyde aqueous solution with a mass concentration of 10% -40% and a dopamine-modified bioactive inorganic material aqueous solution with a mass concentration of 2% -10%, and is prepared according to the following steps (22-28): (7-10): (4-10) mixing FCB hydrogel obtained by Schiff base reaction in a volume ratio, wherein F is polyether-cationic polymer; c is polyether-p-hydroxybenzaldehyde; b is a dopamine modified bioactive inorganic material.

Preferably, the polyether is one of polyethylene glycol (PEG), polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer (P123) and Pluronic F127.

Preferably, the cationic polymer is one of Polyethyleneimine (PEI), polypropyleneimine (PPI), Polylysine (PLL), epsilon-polylysine (EPL), and Chitosan (Chitosan).

Preferably, the bioactive inorganic material is one of Bioactive Glass (BGN), Hydroxyapatite (HAP), Calcium Phosphate (CP), β -tricalcium phosphate (β -TCP), calcium bidirectional phosphate (BCP), Calcium Silicate (CS), and Calcium Carbonate (CC).

A preparation method of self-healing hydrogel for promoting wound healing and tumor treatment comprises the following steps of preparing polyether-p-hydroxybenzaldehyde into aqueous solution with the mass concentration of 10% -40%, preparing polyether-cationic polymer into aqueous solution with the mass concentration of 8% -50%, preparing PDA-bioactive inorganic material into aqueous solution with the mass concentration of 2% -10%, and mixing according to the ratio of (7-10): (22-28): (4-10) taking the three aqueous solutions according to the volume ratio; and then adding the polyether-p-hydroxybenzaldehyde aqueous solution and the PDA-bioactive inorganic material aqueous solution into the polyether-cationic polymer aqueous solution serving as the main structure, uniformly mixing, standing at 37 ℃ for 6-24h, and forming a gel network through Schiff base reaction to obtain the FCB hydrogel.

Preferably, the preparation method of the polyether-p-hydroxybenzaldehyde comprises the following steps:

polyether is reacted with p-toluenesulfonate to prepare polyether-Ots;

polyether-OTs with 4-hydroxybenzaldehyde and potassium carbonate (K)₂CO₃) The reaction produced polyether-Phe-CHO.

Preferably, the preparation method of the polyether-cationic polymer is as follows:

polyether is reacted with p-toluenesulfonate to prepare polyether-Ots;

polyether-OTs are reacted with cationic polymers to prepare polyether-cationic polymer copolymers.

Preferably, the preparation method of the dopamine modified bioactive inorganic material comprises the following steps:

the PDA-bioactive inorganic material is obtained by modifying the surface of the bioactive inorganic material after dopamine is polymerized into poly-dopamine by self.

Compared with the prior art, the invention has the following beneficial technical effects:

the polyether-Phe-CHO and the polyether-cationic polymer are prepared by taking the polyether with better biocompatibility and the cationic polymer with stronger antibacterial property as raw materials; then, dopamine is self-polymerized into poly-dopamine, and then the poly-dopamine is modified on the surface of the bioactive inorganic material to obtain the PDA-bioactive inorganic material. And mixing polyether-Phe-CHO and PDA-bioactive inorganic materials with polyether-cationic polymer, and reacting with Schiff base to obtain FCB hydrogel. The preparation method is simple, no organic solvent residue is generated, the used synthetic method is environment-friendly, the operation is convenient, and the raw material cost is low. Has the following advantages:

(1) the polyether used in the invention has good biocompatibility, and is cheap and easy to obtain.

(2) According to the invention, the cationic polymer and polyether are reacted to prepare polyether-cationic polymer which is used as one of the components of the hydrogel, so that the hydrogel is endowed with excellent antibacterial performance.

(3) The invention utilizes polydopamine to modify the bioactive inorganic material, improves the problem of poor water solubility of the bioactive inorganic material, not only endows the bioactive inorganic material with photo-thermal performance, but also promotes the repair of skin injury.

(4) The FCB hydrogel prepared by the invention can be injected and can be self-healed.

(5) The FCB hydrogel prepared by the method has good photo-thermal property under laser irradiation, and can effectively inhibit the growth of animal tumors.

(6) The solvent used in the present invention is water, and the FCB hydrogel prepared does not contain any organic solvent.

The FCB hydrogel is proved by experimental results to be as follows: the compound has good biocompatibility, good photo-thermal property and strong antibacterial property, can effectively inhibit the growth of animal tumors under the irradiation of laser, and can promote the repair of the skin injury of mice.

Drawings

FIG. 1A shows the FCB1 hydrogel FEPL¹H-NMR nuclear magnetic spectrum;

FIG. 1B is an infrared spectrum of the components of the FCB1 hydrogel.

FIG. 2A is a photograph of the FCB1 hydrogel-forming process;

fig. 2B is the FCB1 hydrogel self-healing process;

fig. 2C is a graph of FCB1 injectable results.

FIG. 3A shows the temperature change of FCB1 and the components after laser irradiation;

fig. 3B is the cytotoxicity results of FCB2 and each component incubated in a375 and C2C12 cells for different times.

Fig. 4 is the results of the FCB3 hydrogel and control group's resistance to escherichia coli (e.coli), staphylococcus aureus (s.aureus) and methicillin-resistant staphylococcus aureus (MRSA).

Figure 5 is a graph of the tumor treatment effect of FCB5 hydrogel and each control group.

Figure 6 is the result of repair of skin lesions in mice with FCB6 hydrogel and each control group.

Detailed Description

The present invention will now be described in further detail with reference to specific examples, which are intended to be illustrative, but not limiting, of the invention.

The invention takes polyether-Phe-CHO, polyether-cationic polymer and PDA-bioactive inorganic material as raw materials to prepare the self-healing hydrogel for promoting wound healing and treating tumors. The polyether-cationic polymer is used as a main structure, has good biocompatibility and strong antibacterial property, and can react with polyether-Phe-CHO and PDA-bioactive inorganic materials through Schiff base to form a gel network. The PDA-bioactive inorganic material modified by polydopamine is used as a photo-thermal preparation to endow hydrogel with photo-thermal performance, can effectively inhibit tumor growth under laser irradiation, and can promote skin wound repair. The multifunctional hydrogel which can be applied to tumor photothermal treatment, skin injury repair and infection resistance is prepared by using polydopamine-modified micro-nano Bioactive Glass (BGN) or biological ceramic material as a photothermal preparation.

Example 1

(1) Preparation of Pluronic F127-p-toluenesulfonate (F127-OTs): 12.3g F127 was dissolved in 70mL of anhydrous dichloromethane, and then 0.83mL of triethylamine and 1.14g of p-toluenesulfonyl chloride were added in that order and reacted at room temperature for 24 h. After the reaction, the organic phase was washed with dilute hydrochloric acid and then with saturated sodium bicarbonate solution, and extracted. The organic phase was then concentrated and precipitated in ether to give F127-OTs (90% yield).

(2) Preparation of F127-p-hydroxybenzaldehyde (F127-Phe-CHO): 2.5g F127-OTs were dissolved in 50mL DMF and 0.11g 4-hydroxybenzaldehyde and 0.12g potassium carbonate (K) were added₂CO₃) The reaction was carried out at 80 ℃ for 72 h. After the reaction was complete, it was cooled to room temperature, 50mL of water was added and extracted with dichloromethane. The organic phase was dried over anhydrous magnesium sulfate and precipitated in ether to give F127-Phe-CHO (90% yield).

(3) Preparation of F127-polylysine Polymer (FEPL): 6.5g F127-OTs and 4.2g EPL were dissolved in 20mL of water and 60mL of DMSO, respectively, and then an aqueous solution of F127-OTs was added to the EPL solution to react at 60 ℃ for 72 hours. After the reaction is finished, dialyzing in distilled water for 3 days by using a dialysis bag with the molecular weight cutoff of 5000, and freeze-drying to obtain the FEPL polymer.

(4) Preparation of polydopamine modified bioactive glass (PDA-BGN): 20mg of BGN was washed three times with Tris and dispersed in 40mL of Tris buffer, and 80mg of Dopamine (DA) was added to the above solution and reacted at room temperature for 1 h. Then, the primary product is washed three times by Tris, and then is dried in vacuum to obtain the PDA-BGN.

(5) Preparation of hydrogel (FCB 1): after 70. mu. L F127-Phe-CHO aqueous solution (20 wt%) and 80. mu.L of PDA-BGN (5 wt%) solution were mixed well, 250. mu.L of FEPL aqueous solution (40 wt%) was added, and after mixing well, the mixture was left at 37 ℃ for 24 hours to obtain FCB hydrogel. Hydrogel FCE1, prepared with 80 μ L of water instead of PDA-BGN solution, served as a control.

The FCB1 hydrogel was injectable, self-healed (fig. 2B), and exhibited strong near infrared light absorption properties (fig. 3A). Under 808nm laser irradiation, the temperature of FCB is raised by 25.9 ℃ within 12 min (FIG. 3A), and the cytotoxicity of FCB1 and each component is low. The hydrogel has strong antibacterial property on gram-negative bacteria (Escherichia coli, E, coli), gram-positive bacteria (Staphylococcus aureus, S, aureus) and methicillin-resistant Staphylococcus aureus (MRSA). The hydrogel was injected into the tumor of xenograft in nude mice, and then the tumor of nude mice was irradiated by laser. After 18 days of treatment, the mice were sacrificed and then the tumors were dissected out and photographed by weighing, and it was observed that the cancer cells were almost completely ablated. FCB1 hydrogel was applied to mouse skin wounds and the skin repair was recorded for each group by taking pictures on

days

0, 3, 7, 10 and 14, respectively. The FCB1 hydrogel was observed to be effective in promoting healing of the skin wound as compared to the control group. Therefore, the FCB1 hydrogel has great application value in anti-infection, cancer treatment and wound healing.

Example 2

(2) Preparation of F127-p-hydroxybenzaldehyde (F127-Phe-CHO): 2.5g F127-OTs were dissolved in 50mL DMF and 0.11g 4-hydroxybenzaldehyde and 0.1g2g Potassium carbonate (K)₂CO₃) The reaction was carried out at 80 ℃ for 72 h. After the reaction was complete, it was cooled to room temperature, 50mL of water was added and extracted with dichloromethane. The organic phase was dried over anhydrous magnesium sulfate and precipitated in ether to give F127-Phe-CHO (90% yield).

(3) Preparation of F127-polyethyleneimine Polymer (FPEI): 6.5g F127-OTs and 0.9g PEI were dissolved in 20mL water and 60mL water, respectively, and the aqueous solution of F127-OTs was added to the PEI solution and reacted at 60 ℃ for 72 h. After the reaction is finished, dialyzing for 3 days in distilled water by using a dialysis bag with the molecular weight cutoff of 3000, and freeze-drying to obtain the FPEI polymer.

(4) Preparation of polydopamine modified calcium phosphate (PDA-CP): 20mg of BGN was washed three times with Tris and dispersed in 40mL of Tris buffer, and then 60mg of Dopamine (DA) was added to the above solution and reacted at room temperature for 1 h. Then, the primary product is washed three times by Tris, and then is dried in vacuum to obtain the PDA-CP.

(5) Preparation of hydrogel (FCB 2): after 80. mu. L F127-Phe-CHO aqueous solution (25 wt%) and 100. mu.L PDA-CP (4 wt%) were mixed, 220. mu.L FPEI aqueous solution (30 wt%) was added, and after mixing, the mixture was left at 37 ℃ for 24 hours to obtain FCB2 hydrogel. Hydrogel FCE2 prepared with 100 μ L of water instead of PDA-CP solution served as a control.

The FCB2 hydrogel can be injected, can be self-healed, and has strong near infrared light absorption property. Under 808nm laser irradiation, FCB2 rapidly increased in temperature, and FCB2 and each component were less cytotoxic (FIG. 3B). The hydrogel has strong antibacterial property on gram-negative bacteria (Escherichia coli, E, coli), gram-positive bacteria (Staphylococcus aureus, S, aureus) and methicillin-resistant Staphylococcus aureus (MRSA). The hydrogel was injected into the tumor of xenograft in nude mice, and then the tumor of nude mice was irradiated by laser. After 18 days of treatment, the mice were sacrificed and then the tumors were dissected out and photographed by weighing, and it was observed that the cancer cells were almost completely ablated. FCB2 hydrogel was applied to mouse skin wounds and the skin repair was recorded for each group by taking pictures on

days

0, 3, 7, 10 and 14, respectively. The FCB2 hydrogel was observed to be effective in promoting healing of the skin wound as compared to the control group. Therefore, the FCB2 hydrogel has great application value in anti-infection, cancer treatment and wound healing.

Example 3

(1) Preparation of P123-P-toluenesulfonate (P123-OTs): 5.8g P123 was dissolved in 70mL of anhydrous dichloromethane, and then 0.56mL of triethylamine and 0.76g of p-toluenesulfonyl chloride were added in that order and reacted at room temperature for 24 hours. After the reaction, the organic phase was washed with dilute hydrochloric acid and then with saturated sodium bicarbonate solution, and extracted. The organic phase was then concentrated and precipitated in ether to give P123-OTs (87% yield).

(2) Preparation of P123-P-hydroxybenzaldehyde (P123-Phe-CHO): 2g P123-OTs were dissolved in 40mL DMF and 0.1g 4-hydroxybenzaldehyde and 0.11g potassium carbonate (K) were added₂CO₃) The reaction was carried out at 80 ℃ for 72 h. After the reaction was complete, it was cooled to room temperature, 40mL of water was added and extracted with dichloromethane. The organic phase was dried over anhydrous magnesium sulfate and precipitated in ether to give P123-Phe-CHO (91% yield).

(3) Preparation of P123-Polylysine Polymer (PPLL): 2.9g P123-OTs and 3.5g PLL were dissolved in 40mL water and 50mL water, respectively, and then an aqueous solution of P123-OTs was added to the PLL solution and reacted at 60 ℃ for 72 h. After the reaction is finished, dialyzing the mixture in distilled water for 3 days by using a dialysis bag with the molecular weight cutoff of 14000, and freeze-drying to obtain the PPLL polymer.

(4) Preparation of Polydopamine modified hydroxyapatite (PDA-HAP): 20mg of HAP was washed three times with Tris and dispersed in 40mL of Tris buffer, and then 100mg of Dopamine (DA) was added to the above solution to react at room temperature for 1 h. Then, the primary product is washed three times with Tris, and then dried in vacuum to obtain PDA-HAP.

(5) Preparation of hydrogel (FCB 3): mu.l of an aqueous solution (10 wt%) of 123-Phe-CHO L P123 and 40. mu.l of a PDA-HAP (8 wt%) solution were mixed, added to 280. mu.l of an aqueous solution (40 wt%) of PPLL, mixed and left at 37 ℃ for 24 hours to obtain FCB3 hydrogel. Hydrogel FCE3 prepared with 40. mu.L of water instead of PDA-HAP solution served as a control.

The FCB3 hydrogel was injectable, self-healed, and exhibited strong near infrared light absorption properties (fig. 3A). Under 808nm laser irradiation, the FCB3 can rapidly increase the temperature, and the cytotoxicity of FCB3 and each component is low. The hydrogel has strong antibacterial property to gram-negative bacteria (Escherichia coli, E, coli), gram-positive bacteria (Staphylococcus aureus, S, aureus) and methicillin-resistant Staphylococcus aureus (MRSA) (FIG. 4). The hydrogel was injected into the tumor of xenograft in nude mice, and then the tumor of nude mice was irradiated by laser. After 18 days of treatment, the mice were sacrificed and then the tumors were dissected out and photographed by weighing, and it was observed that the cancer cells were almost completely ablated. FCB3 hydrogel was applied to mouse skin wounds and the skin repair was recorded for each group by taking pictures on

days

0, 3, 7, 10 and 14, respectively. The FCB3 hydrogel was observed to be effective in promoting healing of the skin wound as compared to the control group. Therefore, the FCB3 hydrogel has great application value in anti-infection, cancer treatment and wound healing.

Example 4

(3) Preparation of P123-Polypropylene imine Polymer (PPPI): 2.9g P123-OTs and 1.2g PPI were dissolved in 40mL of water and 50mL of water, respectively, and then the aqueous solution of P123-OTs was added to the PLL solution and reacted at 60 ℃ for 72 hours. After the reaction is finished, dialyzing the mixture in distilled water for 3 days by using a dialysis bag with the molecular weight cutoff of 3000, and freeze-drying to obtain the PPPI polymer.

(4) Preparation of polydopamine modified β -tricalcium phosphate (β -TCP) (PDA- β -TCP): 20mg of β -TCP was washed three times with Tris and dispersed in 40mL of Tris buffer, and then 120mg of Dopamine (DA) was added to the above solution to react at room temperature for 1 h. Then, the primary product is washed three times by Tris, and is dried in vacuum to obtain the PDA-beta-TCP.

(5) Preparation of hydrogel (FCB 4): mu.l of an aqueous solution (30 wt%) of 123-Phe-CHO L P123 and 40. mu.l of a PDA-beta-TCP (6 wt%) solution were mixed, added to 260. mu.l of an aqueous solution (35 wt%) of PPPI, mixed and left at 37 ℃ for 6 hours to obtain an FCB4 hydrogel. Hydrogel FCE4, prepared with 40 μ L of water instead of PDA- β -TCP solution, served as a control.

The FCB4 hydrogel can be injected, can be self-healed, and has strong near infrared light absorption property. Under 808nm laser irradiation, the FCB4 can rapidly increase the temperature, and the cytotoxicity of FCB4 and each component is low. The hydrogel has strong antibacterial property on gram-negative bacteria (Escherichia coli, E, coli), gram-positive bacteria (Staphylococcus aureus, S, aureus) and methicillin-resistant Staphylococcus aureus (MRSA). The hydrogel was injected into the tumor of xenograft in nude mice, and then the tumor of nude mice was irradiated by laser. After 18 days of treatment, the mice were sacrificed and then the tumors were dissected out and photographed by weighing, and it was observed that the cancer cells were almost completely ablated. FCB4 hydrogel was applied to mouse skin wounds and the skin repair was recorded for each group by taking pictures on

days

0, 3, 7, 10 and 14, respectively. The FCB4 hydrogel was observed to be effective in promoting healing of the skin wound as compared to the control group. Therefore, the FCB4 hydrogel has great application value in anti-infection, cancer treatment and wound healing.

Example 5

(1) Preparation of PEG-p-toluenesulfonate (PEG-OTs): 5g of PEG was dissolved in 70mL of anhydrous dichloromethane, and then 0.56mL of triethylamine and 0.76g of p-toluenesulfonyl chloride were sequentially added and reacted at room temperature for 24 hours. After the reaction, the organic phase was washed with dilute hydrochloric acid and then with saturated sodium bicarbonate solution, and extracted. The organic phase was then concentrated and precipitated in ether to yield PEG-OTs (89% yield).

(2) Preparation of PEG-p-hydroxybenzaldehyde (PEG-Phe-CHO): 2.5g PEG-OTs were dissolved in 50mL DMF, then 0.15g 4-hydroxybenzaldehyde and 0.17g carbon were addedPotassium salt (K)₂CO₃) The reaction was carried out at 80 ℃ for 72 h. After the reaction was complete, it was cooled to room temperature, 50mL of water was added and extracted with dichloromethane. The organic phase was dried over anhydrous magnesium sulfate and precipitated in ether to yield PEG-Phe-CHO (90% yield).

(3) Preparation of PEG-Polyethyleneimine Polymer (PPEI): 2.5g of PEG-OTs and 0.9g of PEI were dissolved in 50mL of water and 20mL of water, respectively, and then the aqueous PEG-OTs solution was added to the PEI solution and reacted at 60 ℃ for 72 hours. After the reaction is finished, dialyzing the mixture in distilled water for 3 days by using a dialysis bag with the molecular weight cutoff of 5000, and freeze-drying to obtain the PPEI polymer.

(4) Preparation of polydopamine-modified tricalcium silicate (CS) (PDA-CS): 20mg of CS was washed three times with Tris and dispersed in 40mL of Tris buffer, and then 100mg of Dopamine (DA) was added to the above solution to react at room temperature for 1 h. Then, the primary product is washed three times by Tris, and then is dried in vacuum to obtain the PDA-CS.

(5) Preparation of hydrogel (FCB 5): after mixing 100. mu.L of PEG-Phe-CHO aqueous solution (25 wt%) and 40. mu.L of PDA-CS (7 wt%), the mixture was added to 260. mu.L of PPEI aqueous solution (50 wt%), and the mixture was left at 37 ℃ for 11 hours to obtain FCB5 hydrogel. Hydrogel FCE5 prepared with 40 μ L of water instead of PDA-CS solution served as a control.

The FCB5 hydrogel can be injected, can be self-healed, and has strong near infrared light absorption property. Under 808nm laser irradiation, the FCB5 can rapidly increase the temperature, and the cytotoxicity of FCB5 and each component is low. The hydrogel has strong antibacterial property on gram-negative bacteria (Escherichia coli, E, coli), gram-positive bacteria (Staphylococcus aureus, S, aureus) and methicillin-resistant Staphylococcus aureus (MRSA). The hydrogel was injected into the tumor of xenograft in nude mice, and then the tumor of nude mice was irradiated by laser. After 18 days of treatment, the mice were sacrificed and then the tumors were dissected out and photographed by weighing, and it was observed that the cancer cells were almost completely ablated (fig. 5). FCB5 hydrogel was applied to mouse skin wounds and the skin repair was recorded for each group by taking pictures on

days

0, 3, 7, 10 and 14, respectively. The FCB5 hydrogel was observed to be effective in promoting healing of the skin wound as compared to the control group. Therefore, the FCB5 hydrogel has great application value in anti-infection, cancer treatment and wound healing.

Example 6

(2) Preparation of PEG-p-hydroxybenzaldehyde (PEG-Phe-CHO): 2.5g PEG-OTs were dissolved in 50mL DMF, then 0.15g 4-hydroxybenzaldehyde and 0.17g potassium carbonate (K) were added₂CO₃) The reaction was carried out at 80 ℃ for 72 h. After the reaction was complete, it was cooled to room temperature, 50mL of water was added and extracted with dichloromethane. The organic phase was dried over anhydrous magnesium sulfate and precipitated in ether to yield PEG-Phe-CHO (90% yield).

(3) Preparation of PEG-Chitosan Polymer (PCTS): 2.5g PEG-OTs and 1g CTS were dissolved in 50mL water and 20mL diluted acetic acid solution, respectively, and then the PEG-OTs aqueous solution was added to the CTS solution and reacted at 60 ℃ for 72 hours. After the reaction is finished, dialyzing the mixture in distilled water for 3 days by using a dialysis bag with the molecular weight cutoff of 14000, and freeze-drying to obtain the PCTS polymer.

(4) Preparation of polydopamine modified calcium bidirectional phosphate (BCP) (PDA-BCP): 20mg of BCP was dispersed in 40mL of Tris buffer after being washed three times with Tris, and 80mg of Dopamine (DA) was added to the above solution to react at room temperature for 1 h. Then, the primary product is washed three times by Tris, and then is dried in vacuum to obtain the PDA-BCP.

(5) Preparation of hydrogel (FCB 6): mu.L of PEG-Phe-CHO aqueous solution (40 wt%) and 80. mu.L of PDA-BCP (2 wt%) solution were mixed, added to 220. mu.L of PCTS aqueous solution (10 wt%), mixed well and left at 37 ℃ for 16 hours to obtain FCB6 hydrogel. Hydrogel FCE6 prepared with 80 μ L of water instead of PDA-BCP solution served as control.

The FCB6 hydrogel can be injected, can be self-healed, and has strong near infrared light absorption property. Under 808nm laser irradiation, the FCB6 can rapidly increase the temperature, and the cytotoxicity of FCB6 and each component is low. The hydrogel has strong antibacterial property on gram-negative bacteria (Escherichia coli, E, coli), gram-positive bacteria (Staphylococcus aureus, S, aureus) and methicillin-resistant Staphylococcus aureus (MRSA). The hydrogel was injected into the tumor of xenograft in nude mice, and then the tumor of nude mice was irradiated by laser. After 18 days of treatment, the mice were sacrificed and then the tumors were dissected out and photographed by weighing, and it was observed that the cancer cells were almost completely ablated. FCB6 hydrogel was applied to mouse skin wounds and the skin repair was recorded for each group by taking pictures on

days

0, 3, 7, 10 and 14, respectively. FCB6 hydrogel was observed to be effective in promoting healing of skin wounds compared to the control group (figure 6). Therefore, the FCB6 hydrogel has great application value in anti-infection, cancer treatment and wound healing.

Example 7

(3) Preparation of PEG-Chitosan Polymer (PCTS): 2g of PEG-OTs and 0.5g of CTS are dissolved in 50mL of water and 20mL of dilute acetic acid solution, respectively, and then the PEG-OTs aqueous solution is added to the CTS solution to react for 72h at 60 ℃. After the reaction is finished, dialyzing the mixture in distilled water for 3 days by using a dialysis bag with the molecular weight cutoff of 14000, and freeze-drying to obtain the PCTS polymer.

(4) Preparation of polydopamine modified Calcium Carbonate (CC) (PDA-CC): 20mg of CC was washed three times with Tris and dispersed in 40mL of Tris buffer, and then 100mg of Dopamine (DA) was added to the above solution to react at room temperature for 1 h. Then, the primary product is washed three times by Tris, and then is dried in vacuum to obtain the PDA-CC.

(5) Preparation of hydrogel (FCB 7): mu.L of PEG-Phe-CHO aqueous solution (40 wt%) and 40. mu.L of PDA-CC aqueous solution (10 wt%) were mixed, added to 260. mu.L of PCTS aqueous solution (8 wt%), mixed well, and left at 37 ℃ for 20 hours to obtain FCB7 hydrogel. Hydrogel FCE7 prepared with 40 μ L of water instead of PDA-CC solution served as a control.

The FCB7 hydrogel can be injected, can be self-healed, and has strong near infrared light absorption property. Under 808nm laser irradiation, the FCB7 can rapidly increase the temperature, and the cytotoxicity of FCB7 and each component is low. The hydrogel has strong antibacterial property on gram-negative bacteria (Escherichia coli, E, coli), gram-positive bacteria (Staphylococcus aureus, S, aureus) and methicillin-resistant Staphylococcus aureus (MRSA). The hydrogel was injected into the tumor of xenograft in nude mice, and then the tumor of nude mice was irradiated by laser. After 18 days of treatment, the mice were sacrificed and then the tumors were dissected out and photographed by weighing, and it was observed that the cancer cells were almost completely ablated. FCB7 hydrogel was applied to mouse skin wounds and the skin repair was recorded for each group by taking pictures on

days

0, 3, 7, 10 and 14, respectively. The FCB7 hydrogel was observed to be effective in promoting healing of the skin wound as compared to the control group. Therefore, the FCB7 hydrogel has great application value in anti-infection, cancer treatment and wound healing.

The FCB hydrogel prepared by the invention can be injected and self-healed, has better photo-thermal property under laser irradiation, and has lower cytotoxicity. In addition, the FCB hydrogel has strong antibacterial performance on gram-negative bacteria, gram-positive bacteria and methicillin-resistant staphylococcus aureus. Moreover, under laser irradiation, the FCB hydrogel can obviously inhibit the growth of tumors in a melanoma model and can effectively promote the skin injury repair of mice. The following is a detailed analysis in conjunction with experimental data.

FIG. 1A shows the FCB1 hydrogel FEPL¹From the H-NMR nuclear magnetic spectrum, it was found that characteristic peaks ascribed to F127 and EPL were observedShown on the spectrum, indicating that FEPL was successfully synthesized; FIG. 1B is an IR spectrum of the constituents of FCB1 hydrogel, which is 1600cm^-1Is the skeleton vibration on the F127-OTs benzene ring, and the C-H stretching vibration and ether bond absorption peaks of the F127 are respectively positioned at 2876cm^-1And 1099cm^-1，3247cm^-1Is the position of the absorption peak of the amino group on the EPL, indicating the successful synthesis of FEPL. 1687cm^-1Peak extinction of aldehyde group and 1662cm^-1Stretching vibration of the imine bond indicated that the aldehyde group of F127-Phe-CHO and the amino group of FEPL reacted via Schiff base to form an FCB1 hydrogel network.

FIG. 2A is a photograph showing the gelation process of FCB1 hydrogel, wherein FCB1 is in solution state at 4 deg.C, and FCB1 is in gel state after a certain period of time when the temperature is increased to 37 deg.C; fig. 2B shows the self-healing process of FCB1 hydrogel, and as shown in the figure, the pores in the gel gradually become smaller and disappear with time, thus proving that FCB1 can self-heal; fig. 2C shows FCB1 hydrogel injectability results, and experiments show that FCB1 can pass through a 1mL syringe and letters can be written.

FIG. 3A shows the temperature change of FCB1 and its components after laser irradiation, which can show the photothermal properties of FCB1 hydrogel, and it can be seen that after laser irradiation, the temperature of FCB1 hydrogel rises to 40 ℃ after 9 minutes of irradiation, compared with FCE1 and water, while the temperature of FCB1 hydrogel without laser irradiation is basically unchanged; fig. 3B shows the cytotoxicity results of FCB2 and each component incubated in a375 and C2C12 cells for different periods of time, and it can be seen from the figure that in a375 and C2C12 cells, the cytotoxicity of FCB2 and each component is small, and even if the cells are incubated for 48 hours, the cell viability is above 80%, which indicates that the FCB hydrogel prepared by the invention has better biocompatibility.

Fig. 4 shows the antibacterial results of FCB3 hydrogel and a control group on escherichia coli (e.coli), staphylococcus aureus (s.aureus) and methicillin-resistant staphylococcus aureus (MRSA), and it can be seen that FCB3 and FCE3 all show higher antibacterial performance among the three bacteria, and the antibacterial ability thereof reaches more than 99%, indicating that the FCB hydrogel prepared by the present invention has stronger anti-infection ability.

Fig. 5 is a graph showing the tumor treatment effect of FCB5 hydrogel and each control group, and it can be seen that FCB5 hydrogel can significantly inhibit tumor growth under laser irradiation, and compared with PBS group, FCB5 has an anti-tumor efficiency as high as 94%.

FIG. 6 shows the results of repairing the FCB6 hydrogel and the skin lesions of each pair of mice, and it can be seen that the area of the skin defect of each group gradually decreased with the lapse of time. Compared with the control group, the FCB6 group showed better wound repair effect, which indicates that the FCB hydrogel can promote wound healing.

The invention uses PDA modified bioactive inorganic material, polyether-Phe-CHO and polyether-cationic polymer as raw materials to prepare FCB hydrogel for the first time. The method has simple process, convenient operation and low cost of raw materials. The prepared FCB hydrogel can be injected and self-healed, has better photothermal property and lower cytotoxicity, and has stronger antibacterial property on gram-negative bacteria, gram-positive bacteria and methicillin-resistant staphylococcus aureus. In addition, the FCB hydrogel has good photo-thermal property under the irradiation of near-infrared laser, can obviously inhibit the growth of tumors at the animal level, and can effectively promote the repair and healing of skin wounds of mice. Therefore, the FCB hydrogel is expected to become a novel multifunctional preparation capable of simultaneously realizing tumor treatment, wound healing and infection resistance, and has a good application prospect in clinical treatment of skin cancer.

The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.

Claims

1. A self-healing hydrogel for promoting wound healing and tumor treatment is characterized in that the self-healing hydrogel is a polyether-cationic polymer aqueous solution with a mass concentration of 8% -50%, a polyether-p-hydroxybenzaldehyde aqueous solution with a mass concentration of 10% -40% and a dopamine-modified bioactive inorganic material aqueous solution with a mass concentration of 2% -10%, and is prepared according to the following steps (22-28): (7-10): (4-10) mixing FCB hydrogel obtained by Schiff base reaction in a volume ratio, wherein F is polyether-cationic polymer; c is polyether-p-hydroxybenzaldehyde; b is a dopamine modified bioactive inorganic material;

the polyether is one of a triblock copolymer of polyethylene oxide-polypropylene oxide-polyethylene oxide and Pluronic F127;

the cationic polymer is one of polyethyleneimine, polypropylene imine and polylysine;

the bioactive inorganic material is one of bioactive glass, hydroxyapatite, calcium phosphate and beta-tricalcium phosphate.

2. A self-healing hydrogel according to claim 1, wherein polylysine is epsilon-polylysine.

3. A method for preparing a self-healing hydrogel for promoting wound healing and tumor treatment according to any one of claims 1 to 2, wherein the polyether-p-hydroxybenzaldehyde is prepared into an aqueous solution with a mass concentration of 10% to 40%, the polyether-cationic polymer is prepared into an aqueous solution with a mass concentration of 8% to 50%, the PDA-bioactive inorganic material is prepared into an aqueous solution with a mass concentration of 2% to 10%, and the following steps are performed: (22-28): (4-10) taking the three aqueous solutions according to the volume ratio; and then adding the polyether-p-hydroxybenzaldehyde aqueous solution and the PDA-bioactive inorganic material aqueous solution into the polyether-cationic polymer aqueous solution serving as the main structure, uniformly mixing, standing at 37 ℃ for 6-24h, and forming a gel network through Schiff base reaction to obtain the FCB hydrogel.

4. A method for preparing a self-healing hydrogel according to claim 3, wherein the polyether-p-hydroxybenzaldehyde is prepared by the following steps:

polyether is reacted with p-toluenesulfonate to prepare polyether-Ots;

polyether-Phe-CHO is prepared by reacting polyether-OTs with 4-hydroxybenzaldehyde and potassium carbonate.

5. A method for preparing a self-healing hydrogel according to claim 3, wherein the polyether-cationic polymer is prepared by the following steps:

polyether is reacted with p-toluenesulfonate to prepare polyether-Ots;

6. A method for preparing a self-healing hydrogel according to claim 3, wherein the dopamine-modified bioactive inorganic material is prepared by the following steps: